Hybrid cucumber &#39;jurassic&#39;

ABSTRACT

A new hybrid cucumber designated ‘Jurassic’ is described. ‘Jurassic’ is an oriental-type slicer cucumber that also has intermediate resistance to cucumber mosaic virus and cucumber vein yellowing virus.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding. Inparticular, this invention relates to a new and distinctive cucumber,Cucumis sativus, hybrid designated ‘Jurassic’.

BACKGROUND OF THE INVENTION

Cultivated forms of cucumber belong to the highly polymorphic speciesCucumis sativus L. that is grown for its edible fruit. As a crop,cucumbers are grown commercially wherever environmental conditionspermit the production of an economically viable yield. They can be handor mechanically harvested. Cucumbers that are grown for fresh market,also called slicers, are generally hand harvested. Those that are to beprocessed are called picklers and may be hand or mechanically harvested.They are produced on trailing or climbing vines. On healthy plants thereis a canopy of large, regular, three-lobed leaves in an alternatearrangement. Pickling cucumbers grown in the United States have usuallyblunt and angular fruits. They are white-spined and most possess darkgreen or medium dark green exterior color. Most slicers have slightlyrounded ends and taper slightly from the stem to blossom end, althoughcylindrical-shaped fruits with blocky or even rounded ends are alsoavailable.

Many changes that occurred with the domestication of the cucumber relateto fruit morphology, with a specialization in fruit shape and size.Slicing cucumbers are frequently sold in lengths from 15 cm to 25 cm anddiameter varies from 2.5 cm to nearly 7 cm. In the United States, theprincipal slicer cucumber growing regions are Georgia, Florida,Michigan, California and North Carolina with nearly 42,000 acres out ofa US total acreage of 57,500 acres. The main states that produceprocessing cucumbers are Michigan, North Carolina and Texas. Freshcucumbers are available in the United States mainly from spring to fall.Cucumbers are consumed in many forms, generally processed for picklingtypes and as fresh market product for slicers. Although slicingcultivars may be processed, they generally are not acceptablesubstitutes for the pickling cucumbers.

Cucumis sativus is a member of the family Cucurbitaceae. TheCucurbitaceae is a family of about 90 genera and 700 to 760 species,mostly of the tropics. The family includes melons, pumpkins, squashes,gourds, watermelon, loofah and many weeds. The genus Cucumis, to whichthe cucumber and several melons belong, includes about 70 species. Thecucumber is believed to be native to India or Southern Asia and has beencultivated there for about 3000 years.

Cucumber is distinct from other Cucumis species in that it has sevenpairs of chromosomes (2n=2x=14) whereas most others have twelve pairs ormultiples of twelve. Pollination techniques for controlled crosses incucumbers are easy to conduct. If bees and natural pollen vectors can beexcluded, the breeder need not be concerned about preventing selfing orother pollen contamination because of the diclinous nature of cucumbersand the stickiness or adherence of pollen to its source flower. There isno wind dissemination of pollen. Pistillate flowers are receptive in themorning or up to midday on the day they open. Cucumbers have a broadrange of floral morphologies, from staminate, pistillate tohermaphrodite flowers, yielding several types of sex expression.

Cucumber is an important and valuable field crop. Thus, there is acontinued need for new hybrid cucumbers. In particular, there is a needfor improved oriental slicer hybrid cucumbers that are stable, highyielding and agronomically sound.

SUMMARY OF THE INVENTION

In order to meet these needs, the present invention is directed toimproved hybrid cucumbers. In one embodiment, the present invention isdirected to a hybrid cucumber, Cucumis sativus, seed designated as‘Jurassic’ having ATCC Accession Number X1. In one embodiment, thepresent invention is directed to a Cucumis sativus cucumber plant andparts isolated therefrom produced by growing ‘Jurassic’ cucumber seed.In another embodiment, the present invention is directed to a Cucumissativus plant and parts isolated therefrom having all the physiologicaland morphological characteristics of a Cucumis sativus plant produced bygrowing ‘Jurassic’ cucumber seed having ATCC Accession Number X1. Instill another embodiment, the present invention is directed to an F₁hybrid Cucumis sativus cucumber seed, plants grown from the seed, andfruit isolated therefrom having ‘Jurassic’ as a parent, where ‘Jurassic’is grown from ‘Jurassic’ cucumber seed having ATCC Accession Number X1.

Cucumber plant parts include cucumber leaves, ovules, pollen, seeds,cucumber fruits, parts of cucumber fruits, flowers, cells, and the like.In another embodiment, the present invention is further directed tocucumber leaves, ovules, pollen, seeds, cucumber fruits, parts ofcucumber fruits, and/or flowers isolated from ‘Jurassic’ cucumberplants. In certain embodiments, the present invention is furtherdirected to pollen or ovules isolated from ‘Jurassic’ cucumber plants.In another embodiment, the present invention is further directed toprotoplasts produced from ‘Jurassic’ cucumber plants. In anotherembodiment, the present invention is further directed to tissue cultureof ‘Jurassic’ cucumber plants, and to cucumber plants regenerated fromthe tissue culture, where the plant has all of the morphological andphysiological characteristics of ‘Jurassic’ cucumber. In certainembodiments, tissue culture of ‘Jurassic’ cucumber plants is producedfrom a plant part selected from leaf, anther, pistil, stem, petiole,root, root tip, fruit, seed, flower, cotyledon, hypocotyl, embryo andmeristematic cell.

In yet another embodiment, the present invention is further directed toa method of selecting cucumber plants, by a) growing ‘Jurassic’ cucumberplants where the ‘Jurassic’ plants are grown from cucumber seed havingATCC Accession Number X1 and b) selecting a plant from step a). Inanother embodiment, the present invention is further directed tocucumber plants, plant parts and seeds produced by the cucumber plantswhere the cucumber plants are isolated by the selection method of theinvention.

In another embodiment, the present invention is further directed to amethod of making cucumber seeds by crossing a cucumber plant grown from‘Jurassic’ cucumber seed having ATCC Accession Number X1 with anothercucumber plant, and harvesting seed therefrom. In still anotherembodiment, the present invention is further directed to cucumberplants, cucumber parts from the cucumber plants, and seeds producedtherefrom where the cucumber plant is grown from seed produced by themethod of making cucumber seed of the invention. In some embodiments,the cucumber plant grown from cucumber seed produced by the method ofmaking cucumber seed is a transgenic lettuce plant.

In another embodiment, the present invention is further directed to amethod of making cucumber variety ‘Jurassic’ by selecting seeds from thecross of one ‘Jurassic’ plant with another ‘Jurassic’ plant, a sample of‘Jurassic’ cucumber seed having been deposited under ATCC AccessionNumber X1.

According to the invention, there is provided a hybrid cucumber plantdesignated ‘Jurassic’. This invention thus relates to the seeds ofhybrid cucumber ‘Jurassic’, to the plants of cucumber ‘Jurassic’ and tomethods for producing a cucumber plant produced by crossing hybridcucumber ‘Jurassic’ with itself or another cucumber plant, and tomethods for producing a cucumber plant containing in its geneticmaterial one or more transgenes and to the transgenic cucumber plantsproduced by that method. This invention also relates to methods forproducing other cucumber cultivars or hybrids derived from hybridcucumber ‘Jurassic’ and to the cucumber cultivars and hybrids derived bythe use of those methods. This invention further relates to cucumberseeds and plants produced by crossing hybrid cucumber ‘Jurassic’ withanother cucumber cultivar.

In another embodiment, the present invention is directed to methods forproducing a cucumber plant containing in its genetic material one ormore transgenes and to the transgenic cucumber plant produced by thosemethods.

In another embodiment, the present invention is directed to single geneconverted plants of hybrid cucumber ‘Jurassic’. The single transferredgene may preferably be a dominant or recessive allele. Preferably, thesingle transferred gene will confer such trait as sex determination,herbicide resistance, insect resistance, resistance for bacterial,fungal, or viral disease, improved harvest characteristics, enhancednutritional quality, or improved agronomic quality. The single gene maybe a naturally occurring cucumber gene or a transgene introduced throughgenetic engineering techniques.

In another embodiment, the present invention is directed to methods fordeveloping cucumber plants in a cucumber plant breeding program usingplant breeding techniques including recurrent selection, backcrossing,pedigree breeding, restriction fragment length polymorphism enhancedselection, genetic marker enhanced selection and transformation. Markerloci such as restriction fragment polymorphisms or random amplified DNAhave been published for many years and may be used for selection (See,Pierce et al., HortScience (1990) 25:605-615; Wehner, T., CucurbitGenetics Cooperative Report, (1997) 20: 66-88; and Kennard et al.,Theorical Applied Genetics (1994) 89:217-224). Seeds, cucumber plants,and parts thereof produced by such breeding methods are also part of theinvention.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by reference bystudy of the following descriptions.

DETAILED DESCRIPTION OF THE INVENTION

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possess the traits to meetthe program goals. The selected germplasm is crossed in order torecombine the desired traits and through selection varieties or parentlines are developed. The goal is to combine in a single variety orhybrid an improved combination of desirable traits from the parentalgermplasm. These important traits may include higher yield, fieldperformance, fruit and agronomic quality such as fruit shape and length,resistance to diseases and insects, and tolerance to drought and heat.

Choice of breeding or selection methods can depend on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially (e.g., F₁ hybrid cultivar, purelinecultivar, etc.). For highly heritable traits, a choice of superiorindividual plants evaluated at a single location will be effective,whereas for traits with low heritability, selection should be based onmean values obtained from replicated evaluations of families of relatedplants. Popular selection methods commonly include pedigree selection,modified pedigree selection, mass selection, and recurrent selection.

The complexity of inheritance influences choice of the breeding method.Backcross breeding is used to transfer one or a few favorable genes fora highly heritable trait into a desirable cultivar. This approach hasbeen used extensively for breeding disease-resistant varieties. Variousrecurrent selection techniques are used to improve quantitativelyinherited traits controlled by numerous genes. The use of recurrentselection in self-pollinating crops depends on the ease of pollination,the frequency of successful hybrids from each pollination, and thenumber of hybrid offspring from each successful cross.

Each breeding program may include a periodic, objective evaluation ofthe efficiency of the breeding procedure. Evaluation criteria varydepending on the goal and objectives, and can include gain fromselection per year based on comparisons to an appropriate standard,overall value of the advanced breeding lines, and number of successfulcultivars produced per unit of input (e.g., per year, per dollarexpended, etc.).

Promising advanced breeding lines are thoroughly tested and compared toappropriate standards in environments representative of the commercialtarget area(s) for at least three years. The best lines can then becandidates for new commercial cultivars. Those still deficient in a fewtraits may be used as parents to produce new populations for furtherselection. These processes, which lead to the final step of marketingand distribution, may take from ten to twenty years from the time thefirst cross or selection is made.

One goal of cucumber plant breeding is to develop new, unique, andgenetically superior cucumber cultivars and hybrids. A breeder caninitially select and cross two or more parental lines, followed byrepeated selfing and selection, producing many new genetic combinations.A plant breeder can then select which germplasms to advance to the nextgeneration. These germplasms may then be grown under differentgeographical, climatic, and soil conditions, and further selections canbe made during, and at the end of, the growing season.

The development of commercial cucumber cultivars thus requires thedevelopment of cucumber parental lines, the crossing of these lines, andthe evaluation of the crosses. Pedigree breeding and recurrent selectionbreeding methods may be used to develop cultivars from breedingpopulations. Breeding programs can be used to combine desirable traitsfrom two or more varieties or various broad-based sources into breedingpools from which lines are developed by selfing and selection of desiredphenotypes. The new lines are crossed with other lines and the hybridsfrom these crosses are evaluated to determine which have commercialpotential.

Pedigree breeding is generally used for the improvement ofself-pollinating crops or inbred lines of cross-pollinating crops. Twoparents which possess favorable, complementary traits are crossed toproduce an F₁. An F₂ population is produced by selfing one or severalF₁'s or by intercrossing two F₁'s (sib mating). Selection of the bestindividuals is usually begun in the F₂ population; then, beginning inthe F₃, the best individuals in the best families are selected.Replicated testing of families, or hybrid combinations involvingindividuals of these families, often follows in the F₄ generation toimprove the effectiveness of selection for traits with low heritability.At an advanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines are tested for potentialrelease as new cultivars.

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified or createdby intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued.

Backcross breeding may be been used to transfer genes for a simplyinherited, highly heritable trait into a desirable homozygous cultivaror line that is the recurrent parent. The source of the trait to betransferred is called the donor parent. The resulting plant is expectedto have the attributes of the recurrent parent (e.g., cultivar) and thedesirable trait transferred from the donor parent. After the initialcross, individuals possessing the phenotype of the donor parent areselected and repeatedly crossed (backcrossed) to the recurrent parent.The resulting plant is expected to have the attributes of the recurrentparent (e.g., cultivar) and the desirable trait transferred from thedonor parent.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F₂ to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F₂ individuals. The number of plants in a population declineseach generation due to failure of some seeds to germinate or some plantsto produce at least one seed. As a result, not all of the F₂ plantsoriginally sampled in the population will be represented by a progenywhen generation advance is completed.

In addition to phenotypic observations, the genotype of a plant can alsobe examined. There are many laboratory-based techniques known in the artthat are available for the analysis, comparison and characterization ofplant genotype. Such techniques include, without limitation, IsozymeElectrophoresis, Restriction Fragment Length Polymorphisms (RFLPs),Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily PrimedPolymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting(DAF), Sequence Characterized Amplified Regions (SCARs), AmplifiedFragment Length polymorphisms (AFLPs), Simple Sequence Repeats(SSRs—which are also referred to as Microsatellites), and SingleNucleotide Polymorphisms (SNPs).

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select toward the genome of the recurrent parent and against themarkers of the donor parent. This procedure attempts to minimize theamount of genome from the donor parent that remains in the selectedplants. It can also be used to reduce the number of crosses back to therecurrent parent needed in a backcrossing program. The use of molecularmarkers in the selection process is often called genetic marker enhancedselection or marker-assisted selection. Molecular markers may also beused to identify and exclude certain sources of germplasm as parentalvarieties or ancestors of a plant by providing a means of trackinggenetic profiles through crosses.

Mutation breeding may also be used to introduce new traits into cucumbervarieties. Mutations that occur spontaneously or are artificiallyinduced can be useful sources of variability for a plant breeder. Thegoal of artificial mutagenesis is to increase the rate of mutation for adesired characteristic. Mutation rates can be increased by manydifferent means including temperature, long-term seed storage, tissueculture conditions, radiation (such as X-rays, Gamma rays, neutrons,Beta radiation, or ultraviolet radiation), chemical mutagens (such asbase analogs like 5-bromo-uracil), antibiotics, alkylating agents (suchas sulfur mustards, nitrogen mustards, epoxides, ethyleneamines,sulfates, sulfonates, sulfones, or lactones), azide, hydroxylamine,nitrous acid or acridines. Once a desired trait is observed throughmutagenesis the trait may then be incorporated into existing germplasmby traditional breeding techniques. Details of mutation breeding can befound in Principles of Cultivar Development by Fehr, MacmillanPublishing Company, 1993.

The production of double haploids can also be used for the developmentof homozygous varieties in a breeding program. Double haploids areproduced by the doubling of a set of chromosomes from a heterozygousplant to produce a completely homozygous individual. For example, seeWan et al., Theor. Appl. Genet., 77:889-892, 1989.

Additional non-limiting examples of breeding methods that may be usedinclude, without limitation, those found in Principles of PlantBreeding, John Wiley and Son, pp. 115-161, 1960; Allard, 1960; Simmonds,1979; Sneep et al., 1979; Fehr, 1987; “Carrots and Related VegetableUmbelliferae”, Rubatzky, V. E., et al., 1999.

Definitions

In the description that follows, a number of terms are used. In order toprovide a clear and consistent understanding of the specification andclaims, including the scope to be given such terms, the followingdefinitions are provided:

Allele. The allele is any of one or more alternative forms of a gene,all of which relate to one trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

Androecious plant. A plant having staminate flowers only.

Backcrossing. Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, a firstgeneration hybrid F₁ with one of the parental genotype of the F₁ hybrid.

Blossom end. The blossom end is the distal end of the fruit (the “far”end as measured from the base of the plant) where the flower blossom islocated. The other end of a fruit is the stem end.

Blossom scar. The blossom scar is the small mark left on the distal endof the fruit after the flower falls off.

Blunt ends. Blunt ends are ends of the cucumber fruits that are nottapered or rounded.

Covered cultivation. Any type of cultivation where the plants are notexposed to direct sunlight. The covering includes but is not limited togreenhouses, glasshouses, nethouses, plastic houses and tunnels.

Essentially all the physiological and morphological characteristics. Aplant having essentially all the physiological and morphologicalcharacteristics means a plant having the physiological and morphologicalcharacteristics of the recurrent parent, except for the characteristicsderived from the converted gene.

Gene. As used herein, “gene” refers to a segment of nucleic acid. A genecan be introduced into a genome of a species, whether from a differentspecies or from the same species, using transformation or variousbreeding methods.

Gynoecious plant. A plant having pistillate flowers only.

Indeterminate vine or Indeterminate Growth. Refers to apical meristemproducing an unrestricted number of lateral organs; characteristic ofvegetative apical meristems. (Anatomy of Seed Plants, 2nd Edition, 1977,John Wiley and Sons, page 513). The main stem of the plant continues togrow as long as the plant stays healthy, as opposed to a determinateplant, which at some point in its life cycle will stop growing longer.

Monoecious plant. A plant having separate staminate and pistillateflowers on the same plant.

Open habit. A plant with an “open habit” means a plant with small tomedium sized leaves (same size or smaller than cucumber cultivar‘Briljant’) and reduced vigor of side shoots (same or less vigor ascucumber cultivar ‘Stereo’).

Parthenocarpic. “Parthenocarpic” refers to the ability of fruit todevelop without pollination or fertilization. The fruit are thereforeseedless.

Percent Identity. Percent identity as used herein refers to thecomparison of the homozygous alleles of two cucumber lines, hybrids orvarieties. Percent identity is determined by comparing a statisticallysignificant number of the homozygous alleles of two developed varieties,lines or hybrids. For example, a percent identity of 90% betweencucumber plant 1 and cucumber plant 2 means that the two plants have thesame allele at 90% of their loci.

Percent Similarity. Percent similarity as used herein refers to thecomparison of the homozygous alleles of a cucumber plant such as hybridcucumber ‘Jurassic’ with another plant, and if the homozygous allele ofhybrid cucumber ‘Jurassic’ matches at least one of the alleles from theother plant then they are scored as similar. Percent similarity isdetermined by comparing a statistically significant number of loci andrecording the number of loci with similar alleles as a percentage. Apercent similarity of 90% between hybrid cucumber ‘Jurassic’ and anotherplant means that hybrid cucumber ‘Jurassic’ matches at least one of thealleles of the other plant at 90% of the loci.

Propagate. To “propagate” a plant means to reproduce the plant by meansincluding, but not limited to, seeds, cuttings, divisions, tissueculture, embryo culture or other in vitro method.

Quantitative Trait Loci (QTL). Quantitative trait loci refer to geneticloci that control to some degree numerically representable traits thatare usually continuously distributed.

Regeneration. Regeneration refers to the development of a plant fromtissue culture.

Single gene converted. Single gene converted or conversion plant refersto plants which are developed by a plant breeding technique calledbackcrossing wherein essentially all of the desired morphological andphysiological characteristics of an inbred are recovered in addition tothe single gene transferred into the inbred via the backcrossingtechnique, genetic engineering or mutation.

Transgene. A “transgene” is a gene taken or copied from one organism andinserted into another organism. A transgene may be a gene that isforeign to the receiving organism or it may be a modified version of anative, or endogenous, gene.

Vertical growing system. A “vertical growing system” means a plantgrowing technique in which plants are grown vertical to the ground withthe use of supporting material. The supporting material includes, but isnot limited to, wires or nets.

Overview of the Hybrid ‘Jurassic’

Hybrid cucumber ‘Jurassic’ is an oriental-type slicer cucumber withsuperior characteristics. Hybrid cucumber ‘Jurassic’ is intended forcovered cultivation from spring to autumn. Hybrid cucumber cultivar‘Jurassic’ is susceptible to Podosphaera xanthii, and resistant tocorynespora leaf spot (Corynespora melonis), cucumber mosaic virus(CMV), and cucumber vein yellowing virus (CVYV). Hybrid cucumbercultivar ‘Jurassic’ is susceptible to zucchini yellow mosaic virus(ZYMV).

Hybrid cucumber ‘Jurassic’ produces fruit having a dark green color, anda long and quite thin shape. Fruit of the hybrid cucumber ‘Jurassic’ isvery ribbed on the outside, contains warts and spines, and has a crunchytexture when eaten. Additionally, hybrid cucumber ‘Jurassic’ has highyield potential, and strong vigor.

Additionally, hybrid cucumber ‘Jurassic’ has shown uniformity andstability for the traits, within the limits of environmental influencefor the traits. Hybrid cucumber ‘Jurassic’ has been increased withcontinued observation for uniformity. No variant traits have beenobserved or are expected in ‘Jurassic’.

Objective Description of the Hybrid ‘Jurassic’

Hybrid cucumber ‘Jurassic’ has the following morphologic and othercharacteristics:

Predominant usage: Slicer

Predominant culture: Covered cultivation

Growing season: Summer

Plant vigor: Medium

Plant sex expression: Male and female flowers approximately equallypresent

Young fruit:

-   -   Type of vestiture: Prickles only    -   Color of vesiture: White

Parthenocarpy: Present

Mature fruit:

-   -   Length: Very long (fruits on main stem +/−39-41 cm)    -   Predominant shape of stem end at market stage: Necked    -   Length of neck: Long    -   Ground color of skin at market stage: Green    -   Intensity of ground color of skin: Medium    -   Ribs: Present    -   Warts and spines: Present

Time of development of female flowers: Late

Cotyledon bitterness: Present

Disease/Pest Resistance:

-   -   Cucumber scab (Cladosporium cucumerinum): Resistant    -   Podosphaera xanthii: Susceptible    -   Corynespora leaf spot (Corynespora melonis): Resistant    -   Cucumber mosaic virus (CMV): Resistant    -   Cucumber vein yellowing virus (CVYV): Resistant    -   Zucchini yellow mosaic virus (ZYMV): Susceptible

Further Embodiments

This invention also is directed to methods for producing a cucumberplant by crossing a first parent cucumber plant with a second parentcucumber plant wherein either the first or second parent cucumber plantis a hybrid cucumber plant of ‘Jurassic.’ Further, both first and secondparent cucumber plants can come from the hybrid cucumber ‘Jurassic.’ Allplants produced using hybrid cucumber ‘Jurassic’ as a parent are withinthe scope of this invention, including plants derived from hybridcucumber ‘Jurassic.’

As used herein, the term plant includes plant cells, plant protoplasts,plant cell tissue cultures from which cucumber plants can beregenerated, plant calli, plant clumps and plant cells that are intactin plants or parts of plants, such as embryos, pollen, ovules, flowers,leaves, stems, and the like.

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants to contain and expressforeign genes, or additional, or modified versions of native, orendogenous, genes (perhaps driven by different promoters) in order toalter the traits of a plant in a specific manner. Such foreignadditional and/or modified genes are referred to herein collectively as“transgenes”. Over the last fifteen to twenty years several methods forproducing transgenic plants have been developed, and the presentinvention, in particular embodiments, also relates to transformedversions of the claimed cultivar.

Plant transformation involves the construction of an expression vectorthat will function in plant cells. Such a vector comprises DNAcomprising a gene under control of or operatively linked to a regulatoryelement (for example, a promoter). The expression vector may contain oneor more such operably linked gene/regulatory element combinations. Thevector(s) may be in the form of a plasmid, and can be used alone or incombination with other plasmids, to provide transformed cucumber plants,using transformation methods as described below to incorporatetransgenes into the genetic material of the cucumber plant(s).

Expression Vectors for Cucumber Transformation: Marker Genes

Expression vectors include at least one genetic marker, operably linkedto a regulatory element (a promoter, for example) that allowstransformed cells containing the marker to be either recovered bynegative selection, i.e., inhibiting growth of cells that do not containthe selectable marker gene, or by positive selection, i.e., screeningfor the product encoded by the genetic marker. Many commonly usedselectable marker genes for plant transformation are well known in thetransformation arts, and include, for example, genes that code forenzymes that metabolically detoxify a selective chemical agent which maybe an antibiotic or a herbicide, or genes that encode an altered targetwhich is insensitive to the inhibitor. A few positive selection methodsare also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptll) gene, isolated from transposonTn5, which when placed under the control of plant regulatory signalswhich confers resistance to kanamycin (Fraley et al., Proc. Natl. Acad.Sci. U.S.A., 80:4803 (1983)). Another commonly used selectable markergene is the hygromycin phosphotransferase gene which confers resistanceto the antibiotic hygromycin (Vanden Elzen et al., Plant Mol. Biol.,5:299 (1985)).

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase, aminoglycoside-3′-adenyl transferase,the bleomycin resistance determinant (Hayford et al., Plant Physiol.86:1216 (1988), Jones et al., Mol. Gen. Genet., 210:86 (1987), Svab etal., Plant Mol. Biol. 14:197 (1990), Hille et al., Plant Mol. Biol.7:171 (1986)). Other selectable marker genes confer resistance toherbicides such as glyphosate, glufosinate or bromoxynil (Comai et al.,Nature 317:741-744 (1985), Gordon-Kamm et al., Plant Cell 2:603-618(1990) and Stalker et al., Science 242:419-423 (1988)).

Selectable marker genes for plant transformation that are not ofbacterial origin include, for example, mouse dihydrofolate reductase,plant 5-enolpyruvylshikimate-3-phosphate synthase and plant acetolactatesynthase (Eichholtz et al., Somatic Cell Mol. Genet. 13:67 (1987), Shahet al., Science 233:478 (1986), Charest et al., Plant Cell Rep. 8:643(1990)).

Another class of marker genes for plant transformation requiresscreening of presumptively transformed plant cells rather than directgenetic selection of transformed cells for resistance to a toxicsubstance such as an antibiotic. These genes are particularly useful toquantify or visualize the spatial pattern of expression of a gene inspecific tissues and are frequently referred to as reporter genesbecause they can be fused to a gene or gene regulatory sequence for theinvestigation of gene expression. Commonly used genes for screeningpresumptively transformed cells include .alpha.-glucuronidase (GUS),.alpha.-galactosidase, luciferase and chloramphenicol, acetyltransferase(Jefferson, R. A., Plant Mol. Biol. Rep. 5:387 (1987), Teeri et al.,EMBO J. 8:343 (1989), Koncz et al., Proc. Natl. Acad. Sci U.S.A. 84:131(1987), DeBlock et al., EMBO J. 3:1681 (1984)).

In vivo methods for visualizing GUS activity that do not requiredestruction of plant tissues are available (Molecular Probes publication2908, IMAGENE GREEN, p.1-4 (1993) and Naleway et al., J. Cell Biol.115:151 a (1991)). However, these in vivo methods for visualizing GUSactivity have not proven useful for recovery of transformed cellsbecause of low sensitivity, high fluorescent backgrounds and limitationsassociated with the use of luciferase genes as selectable markers.

More recently, a gene encoding Green Fluorescent Protein (GFP) has beenutilized as a marker for gene expression in prokaryotic and eukaryoticcells (Chalfie et al., Science 263:802 (1994)). GFP and mutants of GFPmay be used as screenable markers.

Expression Vectors for Cucumber Transformation: Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element, for example, a promoter.Several types of promoters are now well known in the transformationarts, as are other regulatory elements that can be used alone or incombination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred”.Promoters which initiate transcription only in certain tissue arereferred to as “tissue-specific”. A “cell type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter which is under environmental control. Examples ofenvironmental conditions that may affect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter which is active under mostenvironmental conditions.

A. Inducible Promoters

An inducible promoter is operably linked to a gene for expression incucumber. Optionally, the inducible promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in cucumber. With an inducible promoter therate of transcription increases in response to an inducing agent.

Any inducible promoter can be used in the instant invention. See Ward etal., Plant Mol. Biol. 22:361-366 (1993). Exemplary inducible promotersinclude, but are not limited to, that from the ACEI system whichresponds to copper (Meft et al., Proc. Natl. Acad. Sci. U.S.A.90:4567-4571 (1993)); In2 gene from maize which responds tobenzenesulfonamide herbicide safeners (Hershey et al., Mol. Gen Genetics227:229-237 (1991) and Gatz et al., Mol. Gen. Genetics 243:32-38 (1994))or Tet repressor from Tn10 (Gatz et al., Mol. Gen. Genetics 227:229-237(1991). A particularly preferred inducible promoter is a promoter thatresponds to an inducing agent to which plants do not normally respond.An exemplary inducible promoter is the inducible promoter from a steroidhormone gene, the transcriptional activity of which is induced by aglucocorticosteroid hormone. Schena et al., Proc. Natl. Acad. Sci.U.S.A. 88:0421 (1991).

B. Constitutive Promoters

A constitutive promoter is operably linked to a gene for expression incucumber or the constitutive promoter is operably linked to a nucleotidesequence encoding a signal sequence which is operably linked to a genefor expression in cucumber.

Many different constitutive promoters can be utilized in the instantinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses such as the 35S promoter from CaMV(Odell et al., Nature 313:810-812 (1985) and the promoters from suchgenes as rice actin (McElroy et al., Plant Cell 2:163-171 (1990));ubiquitin (Christensen et al., Plant Mol. Biol. 12:619-632 (1989) andChristensen et al., Plant Mol. Biol. 18:675-689 (1992)); pEMU (Last etal., Theor. Appl. Genet. 81:581-588 (1991)); MAS (Velten et al., EMBO J.3:2723-2730 (1984)) and maize H3 histone (Lepetit et al., Mol. Gen.Genetics 231:276-285 (1992) and Atanassova et al., Plant Journal 2 (3):291-300 (1992)). The ALS promoter, Xbal/Ncol fragment 5′ to the Brassicanapus ALS3 structural gene (or a nucleotide sequence similarity to saidXbal/Ncol fragment), represents a particularly useful constitutivepromoter. See PCT application WO 96/30530.

C. Tissue-Specific or Tissue-Preferred Promoters

A tissue-specific promoter is operably linked to a gene for expressionin cucumber. Optionally, the tissue-specific promoter is operably linkedto a nucleotide sequence encoding a signal sequence which is operablylinked to a gene for expression in cucumber. Plants transformed with agene of interest operably linked to a tissue-specific promoter producethe protein product of the transgene exclusively, or preferentially, ina specific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in theinstant invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferred promoter,such as that from the phaseolin gene (Murai et al., Science 23:476-482(1983) and Sengupta-Gopalan et al., Proc. Natl. Acad. Sci. U.S.A.82:3320-3324 (1985)); a leaf-specific and light-induced promoter such asthat from cab or rubisco (Simpson et al., EMBO J. 4(11):2723-2729 (1985)and Timko et al., Nature 318:579-582 (1985)); an anther-specificpromoter such as that from LAT52 (Twell et al., Mol. Gen. Genetics217:240-245 (1989)); a pollen-specific promoter such as that from Zm13(Guerrero et al., Mol. Gen. Genetics 244:161-168 (1993)) or amicrospore-preferred promoter such as that from apg (Twell et al., Sex.Plant Reprod. 6:217-224 (1993)).

Signal Sequences for Targeting Proteins to Subcellular Compartments

Transport of protein produced by transgenes to a subcellular compartmentsuch as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall ormitochondrion or for secretion into the apoplast, is accomplished bymeans of operably linking the nucleotide sequence encoding a signalsequence to the 5′ and/or 3′ region of a gene encoding the protein ofinterest. Targeting sequences at the 5′ and/or 3′ end of the structuralgene may determine, during protein synthesis and processing, where theencoded protein is ultimately compartmentalized.

The presence of a signal sequence directs a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample Becker et al., Plant Mol. Biol. 20:49 (1992), Close, P. S.,Master's Thesis, Iowa State University (1993), Knox, C., et al.,“Structure and Organization of Two Divergent Alpha-Amylase Genes fromBarley”, Plant Mol. Biol. 9:3-17 (1987), Lerner et al., Plant Physiol.91:124-129 (1989), Fontes et al., Plant Cell 3:483-496 (1991), Matsuokaet al., Proc. Natl. Acad. Sci. 88:834 (1991), Gould et al., J. Cell.Biol. 108:1657 (1989), Creissen et al., Plant J. 2:129 (1991), Kalderon,et al., A short amino acid sequence able to specify nuclear location,Cell 39:499-509 (1984), Steifel, et al., Expression of a maize cell wallhydroxyproline-rich glycoprotein gene in early leaf and root vasculardifferentiation, Plant Cell 2:785-793 (1990).

Foreign Protein Genes and Agronomic Genes

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem. 114:92-6(1981).

According to a preferred embodiment, the transgenic plant provided forcommercial production of foreign protein is cucumber. In anotherpreferred embodiment, the biomass of interest is seed. For therelatively small number of transgenic plants that show higher levels ofexpression, a genetic map can be generated, primarily via conventionalRFLP, PCR and SSR analysis, which identifies the approximate chromosomallocation of the integrated DNA molecule. For exemplary methodologies inthis regard, see Glick and Thompson, Methods in Plant Molecular Biologyand Biotechnology, CRC Press, Boca Raton 269:284 (1993). Map informationconcerning chromosomal location is useful for proprietary protection ofa subject transgenic plant. If unauthorized propagation is undertakenand crosses made with other germplasm, the map of the integration regioncan be compared to similar maps for suspect plants, to determine if thelatter have a common parentage with the subject plant. Map comparisonswould involve hybridizations, RFLP, PCR, SSR and sequencing, all ofwhich are conventional techniques.

Likewise, by means of the present invention, agronomic genes can beexpressed in transformed plants. More particularly, plants can begenetically engineered to express various phenotypes of agronomicinterest. Exemplary genes implicated in this regard include, but are notlimited to, those categorized below:

1. Genes That Confer Resistance to Pests or Disease and That Encode:

A. Plant disease resistance genes. Plant defenses are often activated byspecific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant line can be transformed with a clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See for example, Shoresh, et al., “Characterization ofa Mitogen-Activated Protein Kinase Gene from Cucumber Required forTrichoderma-Conferred Plant Resistance”, Plant Physiol. November 2006;142(3): 1169-1179 (activation of Trichoderma-induced MAPK (TIPK) gene isnecessary for the plant's Trichoderma-conferred defense againstbacterial pathogens); Jones et al., Science 266:789 (1994) (cloning ofthe tomato Cf-9 gene for resistance to Cladosporium fulvum); Martin etal., Science 262:1432 (1993) (tomato Pto gene for resistance toPseudomonas syringae pv. tomato encodes a protein kinase); Mindrinos etal., Cell 78:1089 (1994) (Arabidopsis RSP2 gene for resistance toPseudomonas syringae).

B. A Bacillus thuringiensis protein, a derivative thereof or a syntheticpolypeptide modeled thereon. See, for example, Geiser et al., Gene48:109 (1986), who disclose the cloning and nucleotide sequence of a Btdelta-endotoxin gene. Moreover, DNA molecules encoding .delta.-endotoxingenes can be purchased from American Type Culture Collection, Manassas,Va., for example, under ATCC Accession Nos. 40098, 67136, 31995 and31998.

C. A lectin. See, for example, the disclosure by Van Damme et al., PlantMolec. Biol. 24:25 (1994), who disclose the nucleotide sequences ofseveral Clivia miniata mannose-binding lectin genes; and Atsushi, etal., “Possible Involvement of Leaf Gibberellins in the Clock-ControlledExpression of XSP30, a Gene Encoding a Xylem Sap Lectin, in CucumberRoots” Plant Physiol. December 2003; 133(4): 1779-1790.

D. A vitamin-binding protein such as avidin. See PCT application US93/06487, the contents of which are hereby incorporated by reference.The application teaches the use of avidin and avidin homologues aslarvicides against insect pests.

E. An enzyme inhibitor, for example, a protease or proteinase inhibitoror an amylase inhibitor. See, for example, Abe et al., J. Biol. Chem.262:16793 (1987) (nucleotide sequence of rice cysteine proteinaseinhibitor), Huub et al., Plant Molec. Biol. 21:985 (1993) (nucleotidesequence of cDNA encoding tobacco proteinase inhibitor I), Sumitani etal., Biosci. Biotech. Biochem. 57:1243 (1993) (nucleotide sequence ofStreptomyces nitrosporeus .alpha.-amylase inhibitor).

F. An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock et al., Nature 344:458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

G. An insect-specific peptide or neuropeptide which, upon expression,disrupts the physiology of the affected pest. For example, see thedisclosures of Regan, J. Biol. Chem. 269:9 (1994) (expression cloningyields DNA coding for insect diuretic hormone receptor), and Pratt etal., Biochem. Biophys. Res. Comm. 163:1243 (1989) (an allostatin isidentified in Diploptera puntata). See also U.S. Pat. No. 5,266,317 toTomalski et al., who disclose genes encoding insect-specific, paralyticneurotoxins.

H. An insect-specific venom produced in nature by a snake, a wasp, etc.For example, see Pang et al., Gene 116:165 (1992), for disclosure ofheterologous expression in plants of a gene coding for a scorpioninsectotoxic peptide.

I. An enzyme responsible for a hyper-accumulation of a monoterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

J. An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTapplication WO 93/02197 in the name of Scott et al., which discloses thenucleotide sequence of a callase gene. DNA molecules which containchitinase-encoding sequences can be obtained, for example, from the ATCCunder Accession Nos. 39637 and 67152. See also Kramer et al., InsectBiochem. Molec. Biol. 23:691 (1993), who teach the nucleotide sequenceof a cDNA encoding tobacco hornworm chitinase, and Kawalleck et al.,Plant Molec. Biol. 21:673 (1993), who provide the nucleotide sequence ofthe parsley ubi4-2 polyubiquitin gene.

K. A molecule that stimulates signal transduction. For example, see thedisclosure by Botella et al., Plant Molec. Biol. 24:757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess etal., Plant Physiol. 104:1467 (1994), who provide the nucleotide sequenceof a maize calmodulin cDNA clone.

L. A hydrophobic moment peptide. See PCT application WO 95/16776(disclosure of peptide derivatives of tachyplesin which inhibit fungalplant pathogens) and PCT application WO 95/18855 (teaches syntheticantimicrobial peptides that confer disease resistance), the respectivecontents of which are hereby incorporated by reference.

M. A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure of Jaynes et al., Plant Sci 89:43 (1993), ofheterologous expression of a cecropin-β, lytic peptide analog to rendertransgenic tobacco plants resistant to Pseudomonas solanacearum.

N. A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy et al., Ann. rev. Phytopathol.28:451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virus,tobacco streak virus, potato virus X, potato virus Y, tobacco etchvirus, tobacco rattle virus and tobacco mosaic virus. Id.

O. An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. SeeTaylor et al., Abstract #497, Seventh Int'l Symposium on MolecularPlant-Microbe Interactions (Edinburgh, Scotland) (1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

P. A virus-specific antibody. See, for example, Tavladoraki et al.,Nature 366:469 (1993), who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

Q. A developmental-arrestive protein produced in nature by a pathogen ora parasite. Thus, fungal endo-α-1,4-D-polygalacturonases facilitatefungal colonization and plant nutrient release by solubilizing plantcell wall homo-.alpha.-1,4-D-galacturonase. See Lamb et al.,Bio/Technology 10:1436 (1992). The cloning and characterization of agene which encodes a bean endopolygalacturonase-inhibiting protein isdescribed by Toubart et al., Plant J. 2:367 (1992).

R. A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann et al., Bio/Technology 10:305 (1992), have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

2. Genes That Confer Resistance to an Herbicide:

A. An herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee et al.,EMBO J. 7:1241 (1988), and Miki et al., Theor. Appl. Genet. 80:449(1990), respectively.

B. Glyphosate (resistance conferred by mutant5-enolpyruvlshikimate-3-phosphate synthase (EPSPS) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase (PAT) and Streptomyceshygroscopicus phosphinothricin-acetyl transferase PAT bar genes), andpyridinoxy or phenoxy proprionic acids and cyclohexones (ACCaseinhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835 toShah, et al., which discloses the nucleotide sequence of a form of EPSPSwhich can confer glyphosate resistance. A DNA molecule encoding a mutantaroA gene can be obtained under ATCC accession number 39256, and thenucleotide sequence of the mutant gene is disclosed in U.S. Pat. No.4,769,061 to Comai. See also Umaballava-Mobapathie in TransgenicResearch. 1999, 8: 1, 33-44 that discloses Lactuca sativa resistant toglufosinate. European patent application No. 0 333 033 to Kumada et al.,and U.S. Pat. No. 4,975,374 to Goodman et al., disclose nucleotidesequences of glutamine synthetase genes which confer resistance toherbicides such as L-phosphinothricin. The nucleotide sequence of aphosphinothricin-acetyl-transferase gene is provided in Europeanapplication No. 0 242 246 to Leemans et al., DeGreef et al.,Bio/Technology 7:61 (1989), describe the production of transgenic plantsthat express chimeric bar genes coding for phosphinothricin acetyltransferase activity. Exemplary of genes conferring resistance tophenoxy proprionic acids and cyclohexones, such as sethoxydim andhaloxyfop are the Acc1-S1, Acc1-S2 and Acc1-S3 genes described byMarshall et al., Theor. Appl. Genet. 83:435 (1992).

C. An herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) and a benzonitrile (nitrilase gene). Przibilla et al.,Plant Cell 3:169 (1991), describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, andDNA molecules containing these genes are available under ATCC AccessionNos. 53435, 67441, and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes et al., Biochem. J.285:173 (1992).

D. Acetohydroxy acid synthase, which has been found to make plants thatexpress this enzyme resistant to multiple types of herbicides, has beenintroduced into a variety of plants. See Hattori et al., Mol. Gen.Genet. 246:419,1995. Other genes that confer tolerance to herbicidesinclude a gene encoding a chimeric protein of rat cytochrome P4507A1 andyeast NADPH-cytochrome P450 oxidoreductase (Shiota et al., PlantPhysiol., 106:17, 1994), genes for glutathione reductase and superoxidedismutase (Aono et al., Plant Cell Physiol. 36:1687, 1995), and genesfor various phosphotransferases (Datta et al., Plant Mol. Biol. 20:619,1992).

E. Protoporphyrinogen oxidase (protox) is necessary for the productionof chlorophyll, which is necessary for all plant survival. The protoxenzyme serves as the target for a variety of herbicidal compounds. Theseherbicides also inhibit growth of all the different species of plantspresent, causing their total destruction. The development of plantscontaining altered protox activity which are resistant to theseherbicides are described in U.S. Pat. Nos. 6,288,306; 6,282,837;5,767,373; and international publication WO 01/12825.

3. Genes That Confer or Contribute to a Value-Added Trait, Such as:

A. Increased iron content of the cucumber, for example by transforming aplant with a soybean ferritin gene as described in Goto et al., ActaHorticulturae. 2000, 521, 101-109.

B. Increased sweetness of the cucumber by transferring a gene coding formonellin that elicits a flavor 100,000 times sweeter than sugar on amolar basis. See Penarrubia et al., Biotechnology. 1992,10: 561-564.Specific regulatory elements involved in Glc repression also have beenidentified in the promoters of the cucumber malate synthase, see Sarah,et al., (1996). “Distinct cis-acting elements direct the germination andsugar responses of the cucumber malate synthase gene.” Mol. Gen. Genet.250: 153-161.

C. Modified fatty acid metabolism, for example, by transforming a plantwith an antisense gene of stearyl-ACP desaturase to increase stearicacid content of the plant. See Knultzon et al., Proc. Natl. Acad. Sci.USA 89:2625 (1992).

D. Modified carbohydrate composition effected, for example, bytransforming plants with a gene coding for an enzyme that alters thebranching pattern of starch. See Shiroza et al., J. Bacteriol. 170:810(1988) (nucleotide sequence of Streptococcus mutantsfructosyltransferase gene), Steinmetz et al., Mol. Gen. Genet. 20:220(1985) (nucleotide sequence of Bacillus subtilis levansucrase gene), Penet al., Bio/Technology 10:292 (1992) (production of transgenic plantsthat express Bacillus lichenifonnis .alpha.-amylase), Elliot et al.,Plant Molec. Biol. 21:515 (1993) (nucleotide sequences of tomatoinvertase genes), SOgaard et al., J. Biol. Chem. 268:22480 (1993)(site-directed mutagenesis of barley .alpha.-amylase gene), and Fisheret al., Plant Physiol. 102:1045 (1993) (maize endosperm starch branchingenzyme II).

4. Genes that Control Male-Sterility

A. Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN-Ac-PPT. See international publication WO 01/29237.

B. Introduction of various stamen-specific promoters. See internationalpublications WO 92/13956 and WO 92/13957.

C. Introduction of the barnase and the barstar genes. See Paul et al.,Plant Mol. Biol. 19:611-622, 1992).

5. Genes that Create a Site for Site Specific DNA Integration.

This includes the introduction of FRT sites that may be used in theFLP/FRT system and/or Lox sites that may be used in the Cre/Loxp system.For example, see Lyznik, et al., Site-Specific Recombination for GeneticEngineering in Plants, Plant Cell Rep (2003) 21:925-932 and WO 99/25821,which are hereby incorporated by reference. Other systems that may beused include the Gin recombinase of phage Mu (Maeser et al., 1991; VickiChandler, The Maize Handbook ch. 118 (Springer-Verlag 1994), the Pinrecombinase of E. coli (Enomoto et al., 1983), and the R/RS system ofthe pSR1 plasmid (Araki et al., 1992).

6. Genes that Affect Abiotic Stress Resistance.

Genes that affect abiotic stress resistance (including but not limitedto flowering, pod and seed development, enhancement of nitrogenutilization efficiency, altered nitrogen responsiveness, droughtresistance or tolerance, cold resistance or tolerance, and saltresistance or tolerance) and increased yield under stress. For example,see: WO 00/73475 where water use efficiency is altered throughalteration of malate; U.S. Pat. No. 5,892,009, U.S. Pat. No. 5,965,705,U.S. Pat. No. 5,929,305, U.S. Pat. No. 5,891,859, U.S. Pat. No.6,417,428, U.S. Pat. No. 6,664,446, U.S. Pat. No. 6,706,866, U.S. Pat.No. 6,717,034, U.S. Pat. No. 6,801,104; International publicationnumbers WO 2000/060089, WO 2001/026459, WO 2001/035725, WO 2001/034726,WO 2001/035727, WO 2001/036444, WO 2001/036597, WO 2001/036598, WO2002/015675, WO 2002/017430, WO 2002/077185, WO 2002/079403, WO2003/013227, WO 2003/013228, WO 2003/014327, WO 2004/031349, WO2004/076638, WO 98/09521, and WO 99/38977 describing genes, includingCBF genes and transcription factors effective in mitigating the negativeeffects of freezing, high salinity, and drought on plants, as well asconferring other positive effects on plant phenotype; U.S. publicationnumber 2004/0148654 and international publication number WO 01/36596where abscisic acid is altered in plants resulting in improved plantphenotype such as increased yield and/or increased tolerance to abioticstress; international publication numbers WO 2000/006341, WO 04/090143;U.S. application Ser. No. 10/817,483 and U.S. Pat. No. 6,992,237 wherecytokinin expression is modified resulting in plants with increasedstress tolerance, such as drought tolerance, and/or increased yield.Also see international publication numbers WO 02/02776, WO 2003/052063;JP2002281975; U.S. Pat. No. 6,084,153; international publication numberWO 01/64898; U.S. Pat. Nos. 6,177,275 and 6,107,547 (enhancement ofnitrogen utilization and altered nitrogen responsiveness). For ethylenealteration, see U.S. publication numbers 2004/0128719, U.S. 2003/0166197and international publication number WO 2000/32761. For planttranscription factors or transcriptional regulators of abiotic stress,see e.g. U.S. publication numbers 2004/0098764 or U.S. 2004/0078852.

Other genes and transcription factors that affect plant growth andagronomic traits such as yield, flowering, plant growth and/or plantstructure, can be introduced or introgressed into plants, see e.g.international publication numbers WO 97/49811 (LHY), WO 98/56918 (ESD4),WO 97/10339; U.S. Pat. No. 6,573,430 (TFL), U.S. Pat. No. 6,713,663(FT); international publication numbers WO 96/14414 (CON), WO 96/38560,WO 01/21822 (VRN1), WO 00/44918 (VRN2), WO 99/49064 (GI), WO 00/46358(FRI), WO 97/29123; U.S. Pat. No. 6,794,560, U.S. Pat. No. 6,307,126(GAI), and international publication numbers WO 99/09174 (D8 and Rht),and WO 2004/076638 and WO 2004/031349 (transcription factors).

Methods for Cucumber Transformation

Numerous methods for plant transformation have been developed, includingbiological and physical, plant transformation protocols. See, forexample, Miki et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, GlickB. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages67-88. In addition, expression vectors and in vitro culture methods forplant cell or tissue transformation and regeneration of plants areavailable. See, for example, Gruber et al., “Vectors for PlantTransformation” in Methods in Plant Molecular Biology and Biotechnology,Glick B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993)pages 89-119.

A. Agrobacterium-Mediated Transformation

One method for introducing an expression vector into plants is based onthe natural transformation system of Agrobacterium. See, for example,Horsch et al., Science 227:1229 (1985), Curtis et al., Journal ofExperimental Botany. 1994, 45: 279, 1441-1449, Torres et al., Plant cellTissue and Organic Culture. 1993, 34: 3, 279-285, Dinant et al.,Molecular Breeding. 1997, 3: 1, 75-86. A. tumefaciens and A. rhizogenesare plant pathogenic soil bacteria which genetically transform plantcells. The Ti and Ri plasmids of A. tumefaciens and A. rhizogenes,respectively, carry genes responsible for genetic transformation of theplant. See, for example, Kado, C. I., Crit. Rev. Plant Sci. 10:1 (1991).Descriptions of Agrobacterium vector systems and methods forAgrobacterium-mediated gene transfer are provided by Gruber et al.,supra, Miki et al., supra, and Moloney et al., Plant Cell Reports 8:238(1989). See also, U.S. Pat. No. 5,591,616 issued Jan. 7, 1997.

B. Direct Gene Transfer

Several methods of plant transformation collectively referred to asdirect gene transfer have been developed as an alternative toAgrobacterium-mediated transformation. A generally applicable method ofplant transformation is microprojectile-mediated transformation whereinDNA is carried on the surface of microprojectiles measuring 1 to 4 μm.The expression vector is introduced into plant tissues with a biolisticdevice that accelerates the microprojectiles to speeds of 300 to 600 m/swhich is sufficient to penetrate plant cell walls and membranes.Russell, D. R., et al. Pl. Cell. Rep. 12(3, January), 165-169 (1993),Aragao, F. J. L., et al. Plant Mol. Biol. 20(2, October), 357-359(1992), Aragao, F. J. L., et al. Pl. Cell. Rep. 12(9, July), 483-490(1993). Aragao, Theor. Appl. Genet. 93: 142-150 (1996), Kim, J.;Minamikawa, T. Plant Science 117: 131-138 (1996), Sanford et al., Part.Sci. Technol. 5:27 (1987), Sanford, J. C., Trends Biotech. 6:299 (1988),Klein et al., Bio/Technology 6:559-563 (1988), Sanford, J. C., PhysiolPlant 7:206 (1990), Klein et al., Biotechnology 10:268 (1992).

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang et al., Bio/Technology 9:996 (1991). Alternatively,liposome and spheroplast fusion have been used to introduce expressionvectors into plants. Deshayes et al., EMBO J., 4:2731 (1985), Christouet al., Proc Natl. Acad. Sci. U.S.A. 84:3962 (1987). Direct uptake ofDNA into protoplasts using CaCl₂ precipitation, polyvinyl alcohol orpoly-L-ornithine has also been reported. Hain et al., Mol. Gen. Genet.199:161 (1985) and Draper et al., Plant Cell Physiol. 23:451 (1982).Electroporation of protoplasts and whole cells and tissues have alsobeen described. Saker, M.; Kuhne, T. Biologia Plantarum 40(4): 507-514(1997/98), Donn et al., In Abstracts of VIIth International Congress onPlant Cell and Tissue Culture IAPTC, A2-38, p. 53 (1990); D'Halluin etal., Plant Cell 4:1495-1505 (1992) and Spencer et al., Plant Mol. Biol.24:51-61 (1994). See also Chupean et al., Biotechnology. 1989, 7: 5,503-508.

Following transformation of cucumber target tissues, expression of theabove-described selectable marker genes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods now well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic line. The transgenic line could then be crossed,with another (non-transformed or transformed) line, in order to producea new transgenic cucumber line. Alternatively, a genetic trait which hasbeen engineered into a particular cucumber cultivar using the foregoingtransformation techniques could be moved into another line usingtraditional backcrossing techniques that are well known in the plantbreeding arts. For example, a backcrossing approach could be used tomove an engineered trait from a public, non-elite inbred line into anelite inbred line, or from an inbred line containing a foreign gene inits genome into an inbred line or lines which do not contain that gene.As used herein, “crossing” can refer to a simple X by Y cross, or theprocess of backcrossing, depending on the context.

Genetic Marker Profile Through SSR and First Generation Progeny

In addition to phenotypic observations, a plant can also be identifiedby its genotype. The genotype of a plant can be characterized through agenetic marker profile which can identify plants of the same variety ora related variety or be used to determine or validate a pedigree.Genetic marker profiles can be obtained by techniques such asRestriction Fragment Length Polymorphisms (RFLPs), Randomly AmplifiedPolymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Amplified Fragment Length Polymorphisms(AFLPs), Simple Sequence Repeats (SSRs) which are also referred to asMicrosatellites, and Single Nucleotide Polymorphisms (SNPs). Forexample, see Cregan et. al, “An Integrated Genetic Linkage Map of theSoybean Genome” Crop Science 39:1464-1490 (1999), and Berry et al.,“Assessing Probability of Ancestry Using Simple Sequence RepeatProfiles: Applications to Maize Inbred Lines and Soybean Varieties”Genetics 165:331-342 (2003), each of which are incorporated by referenceherein in their entirety.

Particular markers used for these purposes are not limited to anyparticular set of markers, but are envisioned to include any type ofmarker and marker profile which provides a means of distinguishingvarieties. One method of comparison is to use only homozygous loci forhybrid cucumber ‘Jurassic.’

Primers and PCR protocols for assaying these and other markers aredisclosed in the Soybase (sponsored by the USDA Agricultural ResearchService and Iowa State University). In addition to being used foridentification of hybrid cucumber ‘Jurassic’ and plant parts and plantcells of variety hybrid cucumber ‘Jurassic,’ the genetic profile may beused to identify a cucumber plant produced through the use of hybridcucumber ‘Jurassic’ or to verify a pedigree for progeny plants producedthrough the use of hybrid cucumber ‘Jurassic.’ The genetic markerprofile is also useful in breeding and developing backcross conversions.

The present invention comprises a cucumber plant characterized bymolecular and physiological data obtained from the representative sampleof said variety deposited with the American Type Culture Collection(ATCC). Further provided by the invention is a cucumber plant formed bythe combination of the disclosed cucumber plant or plant cell withanother cucumber plant or cell and comprising the homozygous alleles ofthe variety.

Means of performing genetic marker profiles using SSR polymorphisms arewell known in the art. SSRs are genetic markers based on polymorphismsin repeated nucleotide sequences, such as microsatellites. A markersystem based on SSRs can be highly informative in linkage analysisrelative to other marker systems in that multiple alleles may bepresent. See for example, Gong, L., et al., “Microsatellites for thegenus Cucurbita and an SSR-based genetic linkage map of Cucurbita pepoL.” Theor Appl Genet. (June 2008) 117(1): 37-48. Another advantage ofthis type of marker is that, through use of flanking primers, detectionof SSRs can be achieved, for example, by the polymerase chain reaction(PCR), thereby eliminating the need for labor-intensive Southernhybridization. The PCR detection is done by use of two oligonucleotideprimers flanking the polymorphic segment of repetitive DNA. Repeatedcycles of heat denaturation of the DNA followed by annealing of theprimers to their complementary sequences at low temperatures, andextension of the annealed primers with DNA polymerase, comprise themajor part of the methodology. Microsatellites for the genus Cucurbitaand an SSR-based genetic linkage map of Cucurbita pepo L.

Following amplification, markers can be scored by electrophoresis of theamplification products. Scoring of marker genotype is based on the sizeof the amplified fragment, which may be measured by the number of basepairs of the fragment. While variation in the primer used or inlaboratory procedures can affect the reported fragment size, relativevalues should remain constant regardless of the specific primer orlaboratory used. When comparing varieties it is preferable if all SSRprofiles are performed in the same lab.

A number of promoters have utility for plant gene expression for anygene of interest including but not limited to selectable markers,scoreable markers, genes for pest tolerance, disease resistance,nutritional enhancements and any other gene of agronomic interest.Examples of constitutive promoters useful for cucumber plant geneexpression include, but are not limited to, the cauliflower mosaic virus(CaMV) P-35S promoter, which confers constitutive, high-level expressionin most plant tissues (see, e.g., Odel et al., 1985), including monocots(see, e.g., Dekeyser et al., 1990; Terada and Shimamoto, 1990); atandemly duplicated version of the CaMV 35S promoter, the enhanced 35Spromoter (P-e35S) the nopaline synthase promoter (An et al., 1988), theoctopine synthase promoter (Fromm et al., 1989); and the figwort mosaicvirus (P-FMV) promoter as described in U.S. Pat. No. 5,378,619 and anenhanced version of the FMV promoter (P-eFMV) where the promotersequence of P-FMV is duplicated in tandem, the cauliflower mosaic virus19S promoter, a sugarcane bacilliform virus promoter, a commelina yellowmottle virus promoter, and other plant DNA virus promoters known toexpress in plant cells.

A variety of plant gene promoters that are regulated in response toenvironmental, hormonal, chemical, and/or developmental signals can beused for expression of an operably linked gene in plant cells, includingpromoters regulated by (1) heat (Callis et al., 1988), (2) light (e.g.,pea rbcS-3A promoter, Kuhlemeier et al., 1989; maize rbcS promoter,Schaffner and Sheen, 1991; or chlorophyll a/b-binding protein promoter,Simpson et al., 1985), (3) hormones, such as abscisic acid (Marcotte etal., 1989), (4) wounding (e.g., wunl, Siebertz et al., 1989); or (5)chemicals such as methyl jasmonate, salicylic acid, or Safener. It mayalso be advantageous to employ organ-specific promoters (e.g., Roshal etal., 1987; Schernthaner et al., 1988; Bustos et al., 1989).

Exemplary nucleic acids which may be introduced to the cucumber lines ofthis invention include, for example, DNA sequences or genes from anotherspecies, or even genes or sequences which originate with or are presentin the same species, but are incorporated into recipient cells bygenetic engineering methods rather than classical reproduction orbreeding techniques. However, the term “exogenous” is also intended torefer to genes that are not normally present in the cell beingtransformed, or perhaps simply not present in the form, structure, etc.,as found in the transforming DNA segment or gene, or genes which arenormally present and that one desires to express in a manner thatdiffers from the natural expression pattern, e.g., to over-express.Thus, the term “exogenous” gene or DNA is intended to refer to any geneor DNA segment that is introduced into a recipient cell, regardless ofwhether a similar gene may already be present in such a cell. The typeof DNA included in the exogenous DNA can include DNA which is alreadypresent in the plant cell, DNA from another plant, DNA from a differentorganism, or a DNA generated externally, such as a DNA sequencecontaining an antisense message of a gene, or a DNA sequence encoding asynthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and couldpotentially be introduced into a cucumber plant according to theinvention. Non-limiting examples of particular genes and correspondingphenotypes one may choose to introduce into a cucumber plant include oneor more genes for insect tolerance, such as a Bacillus thuringiensis(B.t.) gene, pest tolerance such as genes for fungal disease control,herbicide tolerance such as genes conferring glyphosate tolerance, andgenes for quality improvements such as yield, nutritional enhancements,environmental or stress tolerances, or any desirable changes in plantphysiology, growth, development, morphology or plant product(s). Forexample, structural genes would include any gene that confers insecttolerance including but not limited to a Bacillus insect control proteingene as described in WO 99/31248, herein incorporated by reference inits entirety, U.S. Pat. No. 5,689,052, herein incorporated by referencein its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, hereinincorporated by reference it their entirety. In another embodiment, thestructural gene can confer tolerance to the herbicide glyphosate asconferred by genes including, but not limited to Agrobacterium strainCP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat.No. 5,633,435, herein incorporated by reference in its entirety, orglyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No.5,463,175, herein incorporated by reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes byencoding a non-translatable RNA molecule that causes the targetedinhibition of expression of an endogenous gene, for example viaantisense- or cosuppression-mediated mechanisms (see, for example, Birdet al., 1991). The RNA could also be a catalytic RNA molecule (i.e., aribozyme) engineered to cleave a desired endogenous mRNA product (seefor example, Gibson and Shillito, 1997). Thus, any gene which produces aprotein or mRNA which expresses a phenotype or morphology change ofinterest is useful for the practice of the present invention.

Single-Gene Conversions

When the terms cucumber plant, cultivar, hybrid or cucumber line areused in the context of the present invention, this also includes anysingle gene conversions of that line. The term “single gene convertedplant” as used herein refers to those cucumber plants which aredeveloped by a plant breeding technique called backcrossing whereinessentially all of the desired morphological and physiologicalcharacteristics of a cultivar are recovered in addition to the singlegene transferred into the line via the backcrossing technique.Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the line. The term “backcrossing” asused herein refers to the repeated crossing of a hybrid progeny back toone of the parental cucumber plants for that line, backcrossing 1, 2, 3,4, 5, 6, 7, 8 or more times to the recurrent parent. The parentalcucumber plant which contributes the gene for the desired characteristicis termed the nonrecurrent or donor parent. This terminology refers tothe fact that the nonrecurrent parent is used one time in the backcrossprotocol and therefore does not recur. The parental cucumber plant towhich the gene or genes from the nonrecurrent parent are transferred isknown as the recurrent parent as it is used for several rounds in thebackcrossing protocol (Poehlman & Sleper, 1994; Fehr, 1987). In atypical backcross protocol, the original cultivar of interest (recurrentparent) is crossed to a second line (nonrecurrent parent) that carriesthe single gene of interest to be transferred. The resulting progenyfrom this cross are then crossed again to the recurrent parent and theprocess is repeated until a cucumber plant is obtained whereinessentially all of the desired morphological and physiologicalcharacteristics of the recurrent parent are recovered in the convertedplant, in addition to the single transferred gene from the nonrecurrentparent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalline. To accomplish this, a single gene of the recurrent cultivar ismodified or substituted with the desired gene from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphological,constitution of the original line. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross, one ofthe major purposes is to add some commercially desirable, agronomicallyimportant trait to the plant. The exact backcrossing protocol willdepend on the characteristic or trait being altered to determine anappropriate testing protocol. Although backcrossing methods aresimplified when the characteristic being transferred is a dominantallele, a recessive allele may also be transferred. In this instance itmay be necessary to introduce a test of the progeny to determine if thedesired characteristic has been successfully transferred.

Many single gene traits have been identified that are not regularlyselected for in the development of a new line but that can be improvedby backcrossing techniques. Single gene traits may or may not betransgenic, examples of these traits include but are not limited to,male sterility, modified fatty acid metabolism, modified carbohydratemetabolism, herbicide resistance, resistance for bacterial, fungal, orviral disease, insect resistance, enhanced nutritional quality,industrial usage, yield stability and yield enhancement. These genes aregenerally inherited through the nucleus. Several of these single genetraits are described in U.S. Pat. Nos. 5,777,196, 5,948,957 and5,969,212, the disclosures of which are specifically hereby incorporatedby reference.

Tissue Culture

Further reproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of cucumber andregeneration of plants therefrom is well known and widely published. Forexample, reference may be had to Teng et al., HortScience. 1992, 27:9,1030-1032 Teng et al., HortScience. 1993, 28: 6, 669-1671, Zhang etal., Journal of Genetics and Breeding. 1992, 46: 3, 287-290, Webb etal., Plant Cell Tissue and Organ Culture. 1994, 38: 1, 77-79, Curtis etal., Journal of Experimental Botany. 1994, 45: 279,1441-1449, Nagata etal., Journal for the American Society for Horticultural Science.2000,125: 6, 669-672, and Ibrahim et al., Plant Cell, Tissue and OrganCulture. (1992), 28(2): 139-145. It is clear from the literature thatthe state of the art is such that these methods of obtaining plants areroutinely used and have a very high rate of success. Thus, anotheraspect of this invention is to provide cells which upon growth anddifferentiation produce cucumber plants having the physiological andmorphological characteristics of hybrid cucumber ‘Jurassic.’

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, meristematic cells, andplant cells that can generate tissue culture that are intact in plantsor parts of plants, such as leaves, pollen, embryos, roots, root tips,anthers, pistils, flowers, seeds, petioles, and the like. Means forpreparing and maintaining plant tissue culture are well known in theart. By way of example, a tissue culture comprising organs has been usedto produce regenerated plants. U.S. Pat. Nos. 5,959,185; 5,973,234 and5,977,445 describe certain techniques, the disclosures of which areincorporated herein by reference.

Tissue culture of cucumber can be used for the in vitro regeneration ofcucumber plants. Tissues cultures of various tissues of cucumber andregeneration of plants therefrom are well-known and published. By way ofexample, tissue cultures, some comprising organs to be used to produceregenerated plants, have been described in Burza et al., Plant Breeding.1995,114: 4, 341-345, Cui Hongwen et al., Report Cucurbit GeneticsCooperative. 1999, 22, 5-7, Pellinen, Angewandte Botanik. 1997, 71:3/4,116-118, Kuijpers et al., Plant Cell Tissue and Organ Culture. 1996,46: 1, 81-83, Colijn-Hooymans et al., Plant Cell Tissue and OrganCulture. 1994, 39: 3, 211-217, Lou et al., HortScience. 1994, 29: 8,906-909, Tabei et al., Breeding Science. 1994, 44: 1, 47-51, Sarmanto etal., Plant Cell Tissue and Organ Culture 31:3 185-193 (1992), Raharjo etal., Reports Cucurbits Genetics Cooperative 15, 35-39 (1992),Garcia-Sobo et al., Reports Cucurbits Genetics Cooperative 15, 40-44(1992), Cade et al., Journal of the American Society for HorticulturalScience 115:4 691-696 (1990), Chee et al., HortScience 25:7, 792-793(1990), Kim et al., HortScience 24:4 702 (1989), Punja et al., PlantCell Report 9:2 61-64 (1990). It should also be mentioned that theregeneration of the cucumber after induction of adventitious shoot budson calli derived from cotyledons, has been described in Msikita et al.,Cucurbit Genetics Cooperative Reports, 11: 5-7 (1988), Kim et al., PlantCell Tissue Organ Culture, 12: 67-74 (1988); Wehner et al., HortScience16: 759-760 (1981) had previously described the induction of buds oncotyledons. Cucumber plants could be regenerated by somaticembryogenesis. These somatic embryos developed either in cellsuspensions derived from calli developed from leaf explants Chee et al.,Plant Cell Report 7: 274-277 (1988) or hypocotyls Rajasekaran et al.,Annals of Botany, 52: p. 417-420 (1983), or directly on cotyledonousCade et al., Cucurbit Genetics Cooperative Reports 11:3-4 (1988) or leafcalli Malepszy et al., Pfanzenphysiologie, 111: 273-276 (1983). It isclear from the literature that the state of the art is such that thesemethods of obtaining plants are “conventional” in the sense that theyare routinely used and have a very high rate of success. Thus, anotheraspect of this invention is to provide cells which upon growth anddifferentiation produce cucumber plants having the physiological andmorphological characteristics of hybrid cucumber ‘Jurassic.’

Additional Breeding Methods

This invention also is directed to methods for producing a cucumberplant by crossing a first parent cucumber plant with a second parentcucumber plant wherein the first or second parent cucumber plant is acucumber plant of ‘Jurassic.’ Further, both first and second parentcucumber plants can come from hybrid cucumber ‘Jurassic.’ Thus, any suchmethods using hybrid cucumber ‘Jurassic’ are part of this invention:selfing, backcrosses, hybrid production, crosses to populations, and thelike. All plants produced using hybrid cucumber ‘Jurassic’ as at leastone parent are within the scope of this invention, including thosedeveloped from cultivars derived from hybrid cucumber ‘Jurassic.’Advantageously, this cucumber plant could be used in crosses with other,different, cucumber plants to produce the first generation (F₁) hybridcucumber seeds and plants with superior characteristics. The cultivar ofthe invention can also be used for transformation where exogenous genesare introduced and expressed by the cultivar of the invention. Geneticvariants created either through traditional breeding methods usinghybrid cucumber ‘Jurassic’ or through transformation of hybrid cucumber‘Jurassic’ by any of a number of protocols known to those of skill inthe art are intended to be within the scope of this invention.

The following describes breeding methods that may be used with hybridcucumber ‘Jurassic’ in the development of further cucumber plants. Onesuch embodiment is a method for developing hybrid cucumber ‘Jurassic’progeny cucumber plants in a cucumber plant breeding program comprising:obtaining the cucumber plant, or a part thereof, of hybrid cucumber‘Jurassic,’ utilizing said plant or plant part as a source of breedingmaterial, and selecting a cucumber ‘Jurassic’ progeny plant withmolecular markers in common with hybrid cucumber ‘Jurassic’ and/or withmorphological and/or physiological characteristics selected from thecharacteristics listed in Table 1. Breeding steps that may be used inthe cucumber plant breeding program include pedigree breeding,backcrossing, mutation breeding, and recurrent selection. In conjunctionwith these steps, techniques such as RFLP-enhanced selection, geneticmarker enhanced selection (for example SSR markers) and the making ofdouble haploids may be utilized.

Another method involves producing a population of hybrid cucumber‘Jurassic’ progeny cucumber plants, comprising crossing hybrid cucumber‘Jurassic’ with another cucumber plant, thereby producing a populationof cucumber plants, which, on average, derive 50% of their alleles fromhybrid cucumber ‘Jurassic.’ A plant of this population may be selectedand repeatedly selfed or sibbed with a cucumber plant resulting fromthese successive filial generations. One embodiment of this invention isthe cucumber plant produced by this method and that has obtained atleast 50% of its alleles from hybrid cucumber ‘Jurassic.’

One of ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is nosignificant difference between the two traits expressed by thosevarieties. For example, see Fehr and Walt, Principles of CultivarDevelopment, p. 261-286 (1987). Thus the invention includes hybridcucumber ‘Jurassic’ progeny cucumber plants comprising a combination ofat least two ‘Jurassic’ traits selected from the group consisting ofthose listed in Table 1 or the ‘Jurassic’ combination of traits listedin the Summary of the Invention, so that said progeny cucumber plant isnot significantly different for said traits than cucumber ‘Jurassic’ asdetermined at the 5% significance level when grown in the sameenvironmental conditions and/or may be characterized by percentsimilarity or identity to hybrid cucumber ‘Jurassic’ as determined bySSR markers. Using techniques described herein, molecular markers may beused to identify said progeny plant as a hybrid cucumber ‘Jurassic’progeny plant. Mean trait values may be used to determine whether traitdifferences are significant, and preferably the traits are measured onplants grown under the same environmental conditions. Once such avariety is developed its value is substantial since it is important toadvance the germplasm base as a whole in order to maintain or improvetraits such as yield, disease resistance, pest resistance, and plantperformance in extreme environmental conditions.

Progeny of hybrid cucumber ‘Jurassic’ may also be characterized throughtheir filial relationship with hybrid cucumber ‘Jurassic,’ as forexample, being within a certain number of breeding crosses of hybridcucumber ‘Jurassic.’ A breeding cross is a cross made to introduce newgenetics into the progeny, and is distinguished from a cross, such as aself or a sib cross, made to select among existing genetic alleles. Thelower the number of breeding crosses in the pedigree, the closer therelationship between hybrid cucumber ‘Jurassic’ and its progeny. Forexample, progeny produced by the methods described herein may be within1, 2, 3, 4 or 5 breeding crosses of hybrid cucumber ‘Jurassic.’

As used herein, the term “plant” includes plant cells, plantprotoplasts, plant cell tissue cultures from which cucumber plants canbe regenerated, plant calli, plant clumps and plant cells that areintact in plants or parts of plants, such as leaves, pollen, embryos,cotyledons, hypocotyl, roots, root tips, anthers, pistils, flowers,seeds, stems and the like.

The use of the terms “a,” “an,” and “the,” and similar referents in thecontext of describing the invention (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, butnot limited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. Forexample, if the range 10-15 is disclosed, then 11, 12, 13, and 14 arealso disclosed. All methods described herein can be performed in anysuitable order unless otherwise indicated herein or otherwise clearlycontradicted by context. The use of any and all examples, or exemplarylanguage (e.g., “such as”) provided herein, is intended merely to betterilluminate the invention and does not pose a limitation on the scope ofthe invention unless otherwise claimed. No language in the specificationshould be construed as indicating any non-claimed element as essentialto the practice of the invention.

While a number of exemplary aspects and embodiments have been discussedabove, those of skill in the art will recognize certain modifications,permutations, additions, and sub-combinations thereof. It is thereforeintended that the following appended claims and claims hereafterintroduced are interpreted to include all such modifications,permutations, additions, and sub-combinations as are within their truespirit and scope.

DEPOSIT INFORMATION

A deposit of the hybrid cucumber ‘Jurassic’ is maintained by Enza ZadenUSA, Inc., having an address at 7 Harris Place, Salinas, Calif. 93901,United States. Access to this deposit will be available during thependency of this application to persons determined by the Commissionerof Patents and Trademarks to be entitled thereto under 37 C.F.R. §1.14and 35 U.S.C. §122. Upon allowance of any claims in this application,all restrictions on the availability to the public of the variety willbe irrevocably removed by affording access to a deposit of at least2,500 seeds of the same variety with the American Type CultureCollection, (ATCC), P.O. Box 1549, MANASSAS, Va. 20108 USA.

Applicants have made available to the public without restriction adeposit of at least 2500 seeds of hybrid cucumber ‘Jurassic’ with theAmerican Type Culture Collection (ATCC), P.O. Box 1549, MANASSAS, Va.20108 USA, with a deposit on (DATE) which has been assigned ATCC numberX1.

The deposit will be maintained in the ATCC depository, which is a publicdepository, for a period of 30 years, or 5 years after the most recentrequest, or for the effective life of the patent, whichever is longer,and will be replaced if a deposit becomes nonviable during that period.

1. A hybrid cucumber seed designated as ‘Jurassic’ having ATCC AccessionNumber X1.
 2. A cucumber plant produced by growing the seed of claim 1.3. A plant part from the plant of claim
 2. 4. The plant part of claim 3,wherein said part is a leaf, an ovule, pollen, a seed, a fruit, a cell,or a portion thereof.
 5. A cucumber plant having all the physiologicaland morphological characteristics of the cucumber plant of claim
 2. 6. Aplant part from the plant of claim
 5. 7. The plant part of claim 6,wherein said part is a leaf, an ovule, pollen, a seed, a fruit, a cell,or a portion thereof.
 8. An F₁ hybrid cucumber plant having ‘Jurassic’as a parent where ‘Jurassic’ is grown from the seed of claim
 1. 9.Pollen or an ovule of the plant of claim
 2. 10. A protoplast producedfrom the plant of claim
 2. 11. A tissue culture of the plant of claim 2.12. The tissue culture of claim 11, wherein said tissue culture isproduced from a plant part selected from the group consisting of leaf,anther, pistil, stem, petiole, root, root tip, fruit, seed, flower,cotyledon, hypocotyl, embryo and meristematic cell.
 13. A cucumber plantregenerated from the tissue culture of claim 11, wherein the plant hasall of the morphological and physiological characteristics of a cucumberplant produced by growing seed designated as ‘Jurassic’ having ATCCAccession Number X1.
 14. A method of making cucumber seeds, said methodcomprising crossing the plant of claim 2 with another cucumber plant andharvesting seed therefrom.
 15. A method of making cucumber variety‘Jurassic’, said method comprising selecting seeds from the cross of one‘Jurassic’ plant with another ‘Jurassic’ plant, a sample of ‘Jurassic’cucumber seed having been deposited under ATCC Accession Number X1.